Journal: Scientific Reports
Article Title: KPC-luciferase-expressing cells elicit an anti-tumor immune response in a mouse model of pancreatic cancer
doi: 10.1038/s41598-024-64053-0
Figure Lengend Snippet: KPC-Luc tumors have increased immune cell infiltration when compared to KPC tumors. Immunofluorescence staining of immune cell infiltration in the tumor (red). Nuclei are stained by DAPI (blue). White arrows point to positive stainings. ( A ) Representative images of immunofluorescence staining of KPC and KPC-Luc tumors 9 and 13 days after cell implantation, showing infiltration of CD8 + T cells. ( B ) Quantitative analysis depicted a significantly increased number of CD8 + T cells infiltrated in the tumors of KPC-Luc cells implanted mice, when compared to KPC cells implanted mice, at 13 days of tumor growth, but not at 9 days. Moreover, in the KPC-Luc group, tumor-infiltrated CD8 + T cells increased between days 9 and 13 after cell implantation; n = 3. ( C ) Representative images of immunofluorescence staining of KPC and KPC-Luc tumors 9 and 13 days after cell implantation, showing infiltration of CD68 + cells (macrophages). ( D ) Quantitative analysis showed no significant differences in the number of CD68 + cells infiltrated in the tumor of KPC and KPC-Luc implanted mice at 9 or 13 days of tumor growth; n = 3; Data is presented as mean ± SD. Two-way ANOVA followed by Sidak’s multiple comparisons; *p < 0.01. Scale bars in ( A ) and ( C ) represent 50 µm.
Article Snippet: For this purpose, slides were deparaffinized with xylol and rehydrated with increasing concentrations of EtOH, blocked with fish serum (37527, Invitrogen) for 20 min and incubated overnight with rabbit primary monoclonal antibody against CD8b (1:500 dilution, ab228965, Sigma) or polyclonal antibody against CD68 (1:500 dilution, ab125212, Sigma), followed by 1 h incubation with a goat anti-rabbit secondary antibody conjugated with Alexa Fluor 647 (1:200 dilution, a21244, Invitrogen), and counterstained with DAPI for 30 min for nuclei staining.
Techniques: Immunofluorescence, Staining